腺病毒介导D2S基因联合溴隐亭治疗对GH3细胞生长的抑制作用  

Adenovirus - mediated gene transfer of human D2S combined with bromocriptine inhibits proliferation of GH3 rat anterior pituitary cells

作　　者：刘洁[1] 蒋小龙[1] 陈云祥[1] 苏志鹏[1] 李强[1] 吴近森[1] 吴哲褒[1] 

机构地区：[1]温州医学院附属第一医院神经外科,325000

出　　处：《中华神经外科杂志》2012年第3期295-298,共4页Chinese Journal of Neurosurgery

基　　金：基金项目：国家自然科学基金资助项目（30800347）

摘　　要：目的探讨腺病毒介导多巴胺2型受体短链亚型基因（D2S）联合溴隐亭对大鼠垂体生长激素腺瘤GH3细胞株体外生长活性的影响。方法应用携带人D2S基因及增强型绿色荧光蛋白（EGFP）报告基因的复制缺陷型腺病毒载体pAd—D2S—EGFP感染大鼠GH3细胞，RT—PCR、Western—blot法检测转染后细胞D2SmRNA及蛋白的表达，并采用CCK一8法检测转染D2S基因后联合溴隐亭药物干预治疗，对GH3细胞生长抑制及诱导凋亡作用，运用光镜及荧光显微镜检测联合作用后细胞形态变化。结果GH3细胞转染D2S基因后检测到D2S基因的转录及蛋白表达较对照组增高。腺病毒转染D2S基因联合溴隐亭干预后第2天，体外培养的GH3细胞存活率明显下降（32．76±3．88）％，与单纯加药组的（101．78±2．05）％及空载体组的（91．79±3．80）％相比，差异有统计学意义（P〈0．01）。光镜下及荧光显微镜结合Hoechst核染色观察到细胞形态出现皱缩脱落、死亡、核碎片等改变。结论腺病毒介导D2S基因联合溴隐亭能有效抑制GH3细胞的增殖。Objective To study the effects of adenovirus - mediated D2S gene combined with bromocriptine therapy on proliferation of GH3 rat anterior pituitary cells. Methods GH3 cells was transfeeted with adenovirus containing dopamine 2 receptor short isoform gene （D2S） and confirmed by real time RT - PCR, Western - blot. Cell eytotoxicity assays were performed using a Cell Counting Kit - 8 （ CCK - 8）. Results Expression levels of D2S mRNA and protein increased in GH3 cells transfected with adenovirus containing D2S gene. The percentage of cell survival of GH3 ceils transfeeted with adenovirus containing D2S gene significantly decreased after bromoeriptine treatment, compared with the control group and the pAd - EGFP transfection group （ 32. 76 ± 3.88 ） %, （ 101.78 ± 2. 05 ） %, （ 91.79 ± 3.80 ） %, respectively, P 〈 0. 01. Conclusion Adenovirus - mediated D2S gene combined with bromoeriptine therapy effectively inhibits GH3 cells growth.

关 键 词：腺病毒载体 GH3细胞 基因治疗 溴隐亭 D2S受体 

分 类 号：R73[医药卫生—肿瘤]